Purification and Characterization of Carboxylesterase from the Seeds of Jatropha curcas

Abstract

Carboxylesterases are hydrolases which catalyze the hydrolysis of various types of esters. Carboxylesterase from the seeds of Jatropha curcas has been purified to homogeneity using ammonium sulfate fractionation, CM-cellulose chromatography, Sephadex G-100 chromatography and preparative polyacrylamide gel electrophoresis (PAGE). The homogeneity of the purified enzyme was confirmed by PAGE, iso-electrofocusing and SDS-PAGE. The molecular weight of the purified enzyme was determined by both gel-permeation chromatography on Sephadex G-150 and SDS-PAGE. The molecular weight determined by Sephadex G-150 chromatography and SDS-PAGE both in the presence and absence of 2-mercaptoethanol was 31 kDa. The isoelectric point of the purified enzyme was found to be 8.9. JCSE-I (J. curcas seed esterase-I) was classified as carboxylesterase on the basis of substrate and inhibitor specificity. The K(m) of JCSE-I with 1-naphthyl acetate, 1-naphthyl propionate, 1-naphthyl butyrate and 2-naphthyl acetate as substrates were found to be 0.0,794, 0.0,658, 0.0,567 and 0.1 mM, respectively. The enzyme exhibited an optimum temperature of 45 °C and an optimum pH of 6.5. The enzyme was stable up to 15 min at 65 °C. The enzyme was resistant towards carbamates (carbaryl and eserine sulfate) and sulphydryl inhibitors (p-chloromercuricbenzoate, PCMB) and inhibited by organophosphates (dichlorvos, parathion and phosphamidon).

Comparative proteomic analysis of somatic embryo maturation in Carica papaya L.

“…This protein is considered a biomarker of competent embryogenic cultures in D. carota [95] and Z. mays [96]. Esterase D is also involved in various cellular functions, such as fruit ripening, abscission, cell expansion, stomatal movement, reproduction and the detoxification of xenobiotics [97]. …”

Identification and characterization of a novel carboxylesterase from Phaseolus vulgaris for detection of organophosphate and carbamates pesticides

“…On the other hand, the physiological substrate of the kidney bean esterase is not known and only naphthyl esters were used in the present study. Substrate specificity studies of purified fractions of bean, pea, Jatropha seed and finger millet esterases with phenyl esters, glyceryl esters and naphthyl esters revealed that, in these cases, esterases preferentially hydrolyzed short chain esters. This activity may be important for the hydrolysis of short chain fatty acid esters during the growth and development of the plant …”

“…Additionally, plant esterase can be produced at low cost, with easy extraction and convenient preservation . More importantly, plant esterase from wheat, soybean and jatropha tree has been shown to have a similar sensitivity to AChE.…”

“…[11][12][13][14][15] Additionally, plant esterase can be produced at low cost, with easy extraction and convenient preservation. 11,12 More importantly, plant esterase from wheat, 13,14 soybean 15,16 and jatropha tree 17 has been shown to have a similar sensitivity to AChE. Kidney bean (Phaseolus vulgaris) is a widely distributed and prominent food source for both humans and animals.…”

Partial Characterization of Protease from the Leaves of Jatropha curcas

“…The plant is a common shrub of 3-6 m and the leaves are medicinally used as remedy against rheumatism and jaundice. The seeds are rich sources of oil and the oil obtained from the seeds is used up in the soap industry and also as source of energy (Subramani, Chandrashekharaiah, Swamy, & Murthy, 2012). The plant has attained economical importance due to these applications in areas with extreme climates and soil conditions because of its extraordinary high drought resistance (Gubitz, Lischnig, Stebbing, & Saddler, 1997).…”